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Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica: cloning, characterization, and search for specific inhibitors

Published online by Cambridge University Press:  16 April 2009

A. LOSSANI ,
A. TORTI ,
S. GATTI ,
A. BRUNO ,
R. MASERATI ,
G. E. WRIGHT  and
F. FOCHER
A. LOSSANI
Affiliation:
Istituto di Genetica Molecolare, CNR, Pavia, Italy
A. TORTI
Affiliation:
Istituto di Genetica Molecolare, CNR, Pavia, Italy
S. GATTI
Affiliation:
Laboratorio di Parassitologia, Servizio Virologia, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
A. BRUNO
Affiliation:
Dipartimento di Malattie Infettive, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
R. MASERATI
Affiliation:
Dipartimento di Malattie Infettive, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
G. E. WRIGHT*
Affiliation:
GLSynthesis Inc., Worcester, MA, USA
F. FOCHER
Affiliation:
Istituto di Genetica Molecolare, CNR, Pavia, Italy
*
*Corresponding author: GLSynthesis Inc., One Innovation Drive, Worcester, MA 01605USA. E-mail: [email protected]
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Summary

Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0·27 μm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0·74 and 0·22 mm, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.

Keywords

Entamoebaamoebiasiskinasesinhibitorsdrugs
Type
Research Article
Information
Parasitology , Volume 136 , Issue 6 , May 2009 , pp. 595 - 602
Copyright
Copyright © Cambridge University Press 2009

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